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Multicolor flow cytometric analysis of IL-33R (ST2) expression on Mouse splenocytes. BALB/c Mouse splenic leucocytes were stained with BD Horizon™ Fixable Viability Stain 510 (Cat. No. 564406) followed by staining with FITC Rat Anti-Mouse CD4 antibody (Cat. No. 553047) and with either APC-Cy7 Rat IgG2a, κ Isotype Control (Cat. No. 552770; Left Plot) or APC-Cy7 Rat Anti-Mouse IL-33R (ST2) antibody (Cat. No. 567829/567901; Right Plot) at 0.5 μg/test. The cells were then fixed and permeabilized using the BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574/562725) and stained with PE Rat Anti-Mouse Foxp3 (Cat. No. 560408/560414). The bivariate pseudocolor density plot showing the correlated expression of Foxp3 versus IL-33R (ST2) [or Ig Isotype control staining] was derived from CD4-positive gated events with the forward and side light-scatter characteristics of live cell-discriminated intact lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
BD Pharmingen™ APC-Cy7 Rat Anti-Mouse IL-33R (ST2)
Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
Recommended Assay Procedures
BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cell and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- APC-Cy7 is a tandem fluorochrome composed of Allophycocyanin (APC), which is excited by laser lines between 595 and 647 nm and serves as an energy donor, coupled to the cyanine dye Cy7™, which acts as an energy acceptor and fluoresces at 780 nm. BD Biosciences Pharmingen has maximized the fluorochrome energy transfer in APC-Cy7, thus maximizing its fluorescence emission intensity, minimizing residual emission from APC, and minimizing required electronic compensation in multilaser-laser flow cytometry systems. Note: Although every effort is made to minimize the lot-to-lot variation in residual emission from APC, it is strongly recommended that every lot be tested for differences in the amount of compensation required and that individual compensation controls are run for each APC-Cy7 conjugate.
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- Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Warning: Some APC-Cy7 and PE-Cy7 conjugates show changes in their emission spectrum with prolonged exposure to formaldehyde. If you are unable to analyze fixed samples within four hours, we recommend that you use BD™ Stabilizing Fixative (Cat. No. 338036).
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Companion Products
The U29-93 monoclonal antibody specifically binds to the mouse Interleukin-33 Receptor (IL-33 Receptor, or IL-33R) which is also known as ST2. The IL-33R exists in either a type I transmembrane or soluble glycoprotein form. These IL-33R forms are encoded by the Il1rl1 (Interleukin-1 receptor-like 1) gene which belongs to the IL-1 Receptor family within the Ig superfamily. The IL-33R is expressed by subsets of T cells, including Th2-like cells and some regulatory T cells, as well as some innate lymphocytes, eosinophils, basophils, and mast cells. The IL-33R (also known as the IL-33R alpha subunit, or IL-33Rα) binds IL-33 and complexes with the IL-1R Accessory Protein (IL1RAP) to form a functional signaling receptor complex that can induce the production of T helper type 2 (Th2) cytokines. The soluble IL-33R may function as a decoy receptor which can block the binding of IL-33 to the transmembrane IL-33R. The IL-33R plays roles in inflammation, immunity, and allergy.
Development References (4)
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Cottagiri M, Nyandjo M, Stephens M, et al. In drug-induced, immune-mediated hepatitis, interleukin-33 reduces hepatitis and improves survival independently and as a consequence of FoxP3+ T-cell activity.. Cell Mol Immunol. 2019; 16(8):706-717. (Clone-specific: Flow cytometry). View Reference
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Hayakawa H, Hayakawa M, Kume A, Tominaga S. Soluble ST2 blocks interleukin-33 signaling in allergic airway inflammation.. J Biol Chem. 2007; 282(36):26369-80. (Biology). View Reference
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Matta BM, Lott JM, Mathews LR, et al. IL-33 is an unconventional Alarmin that stimulates IL-2 secretion by dendritic cells to selectively expand IL-33R/ST2+ regulatory T cells.. J Immunol. 2014; 193(8):4010-20. (Biology). View Reference
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Schiering C, Krausgruber T, Chomka A, et al. The alarmin IL-33 promotes regulatory T-cell function in the intestine.. Nature. 2014; 513(7519):564-8. (Biology). View Reference
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.